Our current work focuses primarily on understanding the differences in microtubule organization between muscles of control mice (WT) and of mdx mice, which serve as a model for Duchenne muscular dystrophy (DMD). Normal mouse muscles have a regular grid-like microtubule network, whereas mdx mouse muscles have a disordered, denser network. The software TeDT, developed in the Light Imaging Section for the analysis of microtubule directionality, is an essential tool in the quantitative assessment of such differences in microtubule organization (Liu et al., 2014). Results obtained in previous years demonstrated that the differences in microtubule orientation can be observed as soon as microtubules start growing from the nucleating Golgi elements. Thus muscle microtubules grow as if Golgi elements themselves, or the nucleating molecules anchored to the Golgi elements, had a specific orientation, disturbed in mdx muscles. Conventional and super-resolution microscopy have been used to investigate the orientation of the Golgi elements. Labeling of the Golgi elements with two antibodies, one for the cis-Golgi protein GM130, the other one for the trans-Golgi protein TGN38 allowed us to determine the orientation of each Golgi element. Plotting Golgi directionality with the software TeDT indeed shows differences between WT and mdx mouse muscles, confirming that differences in microtubule organization are sealed at an early stage of their formation. We have also carried out RNA-seq analysis of three different mouse muscles (FDB, EDL, and soleus) at two different ages, 2 and 5 months, from both WT and mdx mice. The goal was a comparison of RNA changes in the mdx mouse muscles compared to human DMD muscles (Khairallah et al. 2012) with special attention to tubulins, the constituents of microtubules, and microtubule-associated proteins. The two ages were selected based on the report in the same paper that microtubules are implicated in the mdx pathology at 5 but not at 2 months of age. The results are still under analysis. They already confirm that several tubulin mRNAs are differentially expressed in mdx compared to WT muscles. In contrast, centrosomal proteins, involved in microtubule nucleation, and MAPs (microtubule-associated proteins) are little affected. Although several mRNAs are differentially expressed in 2 and 5 mo-old mice, this is not the case for tubulin mRNAs. Thus, the difference in response between these ages must be searched in other parts of the pathways affected. After analysis of the results is complete we will further investigate the role of specific molecules hypothesized to play a role in the disorganization of the mdx network.